MicroRNA-378 regulates adipogenic differentiation in bovine intramuscular preadipocytes by targeting CaMKK2.

IF 3.5 4区 生物学 Q2 ENDOCRINOLOGY & METABOLISM
Dongwei Li, Heng Wang, Yongmin Li, Changqing Qu, Yunhai Zhang, Hongyu Liu, Xiaorong Zhang
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引用次数: 4

Abstract

Intramuscular fat, as one of the most important palatability attribute of beef carcase, is the primary determinant of beef quality. The research of adipogenesis mechanism would provide new insight into intramuscular fatty deposition. Here, the role of microRNA-378 was investigated during bovine adipogenic differentiation. It was revealed that miR-378 expression exists variably in bovine major tissue and organs by RT-qPCR. It was predicted that miR-378 targets CaMKK2, as an AMPKα kinase, by DIANA Tools. For better research, primary preadipocytes with stable transfection for up-/down-regulated expression of miR-378 were constructed by lentiviral vectors with GFP gene. The analyses of qPCR showed that PPARγ and adiponectin mRNA levels increased, but C/EBPβ, pref-1 and CaMKK2 mRNA levels decreased during adipogenic differentiation. When miR-378 was overexpressed, preadipocytes proliferation became slower, there are more cellular lipid droplets, and PPARγ and C/EBPβ mRNA levels were higher, but pref-1, adiponectin and CaMKK2 were lower than control groups. Luciferase assay and western blot analysis validated that miR-378 binds the nucleotide sites of the 3'- untranslated region of CaMKK2, which inhibits the mRNA and protein expression of CaMKK2. These findings suggest that miR-378 promotes adipogenic differentiation in bovine intramuscular preadipocytes by targeting CaMKK2 via AMPK signalling pathway.

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MicroRNA-378通过靶向CaMKK2调控牛肌内前脂肪细胞的成脂分化。
肌内脂肪是牛肉胴体最重要的适口性属性之一,是牛肉品质的主要决定因素。脂肪形成机制的研究将为肌内脂肪沉积提供新的认识。本文研究了microRNA-378在牛成脂分化过程中的作用。RT-qPCR结果显示,miR-378在牛主要组织和器官中有不同程度的表达。DIANA Tools预测miR-378作为AMPKα激酶靶向CaMKK2。为了更好地进行研究,我们利用带有GFP基因的慢病毒载体构建了稳定转染miR-378上调/下调表达的原代前脂肪细胞。qPCR分析显示,在成脂分化过程中,PPARγ和脂联素mRNA水平升高,而C/EBPβ、pref-1和CaMKK2 mRNA水平降低。当miR-378过表达时,前脂肪细胞增殖减慢,细胞脂滴增多,PPARγ和C/EBPβ mRNA水平升高,而pref-1、脂联素和CaMKK2水平低于对照组。荧光素酶分析和western blot分析证实miR-378结合CaMKK2 3'-非翻译区核苷酸位点,抑制CaMKK2 mRNA和蛋白的表达。这些发现表明,miR-378通过AMPK信号通路靶向CaMKK2,促进牛肌内前脂肪细胞的成脂分化。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
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来源期刊
Adipocyte
Adipocyte Medicine-Histology
CiteScore
6.50
自引率
3.00%
发文量
46
审稿时长
32 weeks
期刊介绍: Adipocyte recognizes that the adipose tissue is the largest endocrine organ in the body, and explores the link between dysfunctional adipose tissue and the growing number of chronic diseases including diabetes, hypertension, cardiovascular disease and cancer. Historically, the primary function of the adipose tissue was limited to energy storage and thermoregulation. However, a plethora of research over the past 3 decades has recognized the dynamic role of the adipose tissue and its contribution to a variety of physiological processes including reproduction, angiogenesis, apoptosis, inflammation, blood pressure, coagulation, fibrinolysis, immunity and general metabolic homeostasis. The field of Adipose Tissue research has grown tremendously, and Adipocyte is the first international peer-reviewed journal of its kind providing a multi-disciplinary forum for research focusing exclusively on all aspects of adipose tissue physiology and pathophysiology. Adipocyte accepts high-profile submissions in basic, translational and clinical research.
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